Burgess D E, Jerrells T
Infect Immun. 1985 Dec;50(3):893-9. doi: 10.1128/iai.50.3.893-899.1985.
Monoclonal antibodies (MAbs) which are reactive with several antigenically distinct variable antigen types were prepared by immunization with Trypanosoma brucei rhodesiense. Certain MAbs were shown to be specific for members of the genus Trypanosoma and not reactive with Leishmania spp. or Plasmodium falciparum by the indirect immunofluorescence assay. These genus-specific MAbs were used to identify the molecular location of these invariant antigen determinants in whole T. brucei rhodesiense antigen preparations. Two monoclonals reacted with a low-molecular-weight doublet of approximately 22,000 relative molecular weight on Western blots of whole trypanosome antigen preparations separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These determinants did not appear to be on the variable surface glycoprotein molecule and were destroyed by trypsin digestion. Binding studies in which live, DEAE-purified, bloodstream trypanosomes were exposed to invariant antigen-specific MAbs suggested these determinants were accessible on living trypanosomes. Genus-specific MAbs also reacted with determinants present in sera from African trypanosomiasis patients in a dot immunobinding assay but not sera from patients with malaria or leishmaniasis. These results suggest that certain invariant molecules of African trypanosomes are immunogenic, possibly accessible on the trypanosome surface, and may be present as circulating invariant antigen in trypanosomiasis patients.
用罗德西亚布氏锥虫免疫制备了与几种抗原性不同的可变抗原类型发生反应的单克隆抗体(MAb)。通过间接免疫荧光试验表明,某些单克隆抗体对锥虫属成员具有特异性,而与利什曼原虫属或恶性疟原虫无反应。这些属特异性单克隆抗体被用于确定这些不变抗原决定簇在整个罗德西亚布氏锥虫抗原制剂中的分子定位。两种单克隆抗体在经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离的整个锥虫抗原制剂的蛋白质印迹上,与相对分子质量约为22,000的低分子量双峰发生反应。这些决定簇似乎不在可变表面糖蛋白分子上,并且会被胰蛋白酶消化破坏。用活的、经二乙氨基乙基(DEAE)纯化的血流锥虫与不变抗原特异性单克隆抗体进行的结合研究表明,这些决定簇在活锥虫上是可及的。在斑点免疫结合试验中,属特异性单克隆抗体也与非洲锥虫病患者血清中的决定簇发生反应,但与疟疾或利什曼病患者的血清不发生反应。这些结果表明,非洲锥虫的某些不变分子具有免疫原性,可能在锥虫表面是可及的,并且可能作为循环不变抗原存在于锥虫病患者体内。